James W Bogenpohl1, Rory M Weston2, Taylor N Foreman1, Kaitlyn E Kitchen1, Michael F Miles2,3. 1. Department of Molecular Biology and Chemistry, Christopher Newport University, Newport News, Virginia, USA. 2. Department of Pharmacology and Toxicology, Virginia Commonwealth University, Richmond, Virginia, USA. 3. VCU Alcohol Research Center, Virginia Commonwealth University, Richmond, Virginia, USA.
Abstract
BACKGROUND: Chloride intracellular channel 4 (CLIC4) is a multifunctional metamorphic protein for which a growing body of evidence supports a major role in the brain's molecular and behavioral responses to ethanol (EtOH). Although key to understanding the functional biology underlying this role, little is known about the cellular and subcellular expression patterns of CLIC4 in brain and how they are affected by EtOH. METHODS: We used qRT-PCR to assess Clic4 mRNA expression in the medial prefrontal cortex (mPFC) of C57BL/6J mice in the absence and presence of acute EtOH exposure. Two complementary immunohistochemical techniques were employed to assess the subcellular localization of the CLIC4 protein and its pattern of expression across brain cell types in the mPFC in the absence and presence of acute EtOH. RESULTS: Through immunohistochemical and stereological techniques, we show that CLIC4 protein is robustly expressed by oligodendrocytes (most abundant), microglia, and astrocytes, with minimal expression in neurons. Following acute EtOH exposure, we observed a rapid increase in Clic4 mRNA expression in female but not male mice and an overall increase in the number of oligodendrocytes and astrocytes expressing the CLIC4 protein. CONCLUSIONS: These findings suggest that Clic4 functions as an early response gene for acute EtOH in brain, which likely underlies its ability to modulate EtOH behavior. Our results also suggest that the role of CLIC4 in the brain's response to EtOH is mediated through oligodendrocytes.
BACKGROUND: Chloride intracellular channel 4 (CLIC4) is a multifunctional metamorphic protein for which a growing body of evidence supports a major role in the brain's molecular and behavioral responses to ethanol (EtOH). Although key to understanding the functional biology underlying this role, little is known about the cellular and subcellular expression patterns of CLIC4 in brain and how they are affected by EtOH. METHODS: We used qRT-PCR to assess Clic4 mRNA expression in the medial prefrontal cortex (mPFC) of C57BL/6J mice in the absence and presence of acute EtOH exposure. Two complementary immunohistochemical techniques were employed to assess the subcellular localization of the CLIC4 protein and its pattern of expression across brain cell types in the mPFC in the absence and presence of acute EtOH. RESULTS: Through immunohistochemical and stereological techniques, we show that CLIC4 protein is robustly expressed by oligodendrocytes (most abundant), microglia, and astrocytes, with minimal expression in neurons. Following acute EtOH exposure, we observed a rapid increase in Clic4 mRNA expression in female but not male mice and an overall increase in the number of oligodendrocytes and astrocytes expressing the CLIC4 protein. CONCLUSIONS: These findings suggest that Clic4 functions as an early response gene for acute EtOH in brain, which likely underlies its ability to modulate EtOH behavior. Our results also suggest that the role of CLIC4 in the brain's response to EtOH is mediated through oligodendrocytes.
Authors: Ester Fernández-Salas; Kwang S Suh; Vladislav V Speransky; Wendy L Bowers; Joshua M Levy; Tracey Adams; Kamal R Pathak; Lindsay E Edwards; Daniel D Hayes; Christina Cheng; Alasdair C Steven; Wendy C Weinberg; Stuart H Yuspa Journal: Mol Cell Biol Date: 2002-06 Impact factor: 4.272
Authors: Dene R Littler; Stephen J Harrop; W Douglas Fairlie; Louise J Brown; Greg J Pankhurst; Susan Pankhurst; Matthew Z DeMaere; Terence J Campbell; Asne R Bauskin; Raffaella Tonini; Michele Mazzanti; Samuel N Breit; Paul M G Curmi Journal: J Biol Chem Date: 2003-11-12 Impact factor: 5.157
Authors: Kwang S Suh; Michihiro Mutoh; Kunio Nagashima; Ester Fernandez-Salas; Lindsay E Edwards; Daniel D Hayes; John M Crutchley; Keith G Marin; Rebecca A Dumont; Joshua M Levy; Christina Cheng; Susan Garfield; Stuart H Yuspa Journal: J Biol Chem Date: 2003-11-10 Impact factor: 5.157